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1.
Mymensingh Med J ; 33(3): 696-705, 2024 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-38944709

RESUMEN

Major causes of acute insult in Hepatitis B virus related acute on chronic liver failure in the Asian region are reactivation of Hepatitis B virus and super infection with hepatitis A and E virus (ACLF). Anti viral therapy should be started as soon as possible in the ACLF patients at presentation while waiting for confirmation by HBV DNA level. This randomized controlled trial was carried out at the Department of Hepatology, BSMMU, Bangladesh from September 2019 to august 2020 with Hepatitis B virus related ACLF patient. This trial was conducted among twenty seven HBV acute on chronic liver failure patient to compare Child Turcotte pugh (CTP) score, Model for end stage liver disease (MELD) score, Asia Pacific Association for study of Liver (APASL) ACLF Research consortium (AARC) score, survival of the patients and HBV DNA level at 3 months with antiviral therapy between tenofovir alafenamide (25mg) and entecavir (0.5mg) group. CTP score, MELD score and AARC score were significantly (p<0.05) decline from baseline to all subsequent follow-up at 1st (at 7 days), 2nd (at 14 days), 3rd (at 30 days) and 4th (at 90 days) in each group but non significant (p>0.05) difference occurred between two group. All twenty seven patients had detectable HBV DNA level at pre-treatment and all survived patients became undectable at 4th, 90 days follow-up. Total 10 patients (37.07%) were survived at 90 days follow-up, out of them seven patients (70.0%) were in tenofovir alafenamide group and three patients (30.0%) were in entecavir group which was statistically significant (p<0.05) in between two group. Hepatic encephalopathy and hepatorenal syndrome were most common causes of death in both groups. Both drugs tenofovir alafenamide and entecavir significantly improves liver functions but the former one is superior regarding survival.


Asunto(s)
Insuficiencia Hepática Crónica Agudizada , Antivirales , Guanina , Tenofovir , Humanos , Tenofovir/uso terapéutico , Tenofovir/análogos & derivados , Guanina/análogos & derivados , Guanina/uso terapéutico , Antivirales/uso terapéutico , Masculino , Insuficiencia Hepática Crónica Agudizada/tratamiento farmacológico , Insuficiencia Hepática Crónica Agudizada/virología , Insuficiencia Hepática Crónica Agudizada/etiología , Femenino , Adulto , Persona de Mediana Edad , Resultado del Tratamiento , Alanina/análogos & derivados , Alanina/uso terapéutico , Hepatitis B/tratamiento farmacológico , Hepatitis B/complicaciones , Virus de la Hepatitis B/genética , Virus de la Hepatitis B/efectos de los fármacos
2.
Med J Malaysia ; 79(Suppl 1): 134-139, 2024 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-38555898

RESUMEN

INTRODUCTION: Night eating is a very common dietary behaviour among university students. This study aims to investigate the relationship between night eating and BMI, stress, sleep quality and duration of study among university students. MATERIALS AND METHODS: A total of 385 university students including foundation and undergraduate students took part in this study. Self-administered online surveys were used to obtain sociodemographic data, and anthropometry measurements including weight and height, night eating during studying, duration of the study, opinion on eating and academic performance, sleep quality, level of depression, anxiety, and stress of the respondents. Questionnaires were validated and IBM SPSS Statistics Software version 26.0 was used to analyse categorical and continuous variables. RESULTS: The findings showed that there was an association between night eaters and coffee consumption with BMI (p<0.001) and sleep quality (p<0.05). However, there was no association (p>0.05) found between the types of food eaten during night studying and the mean duration of the study. The results showed drinking coffee had an association with depression, anxiety, and stress (p<0.05) among Malaysian university students. CONCLUSION: Coffee consumption was common among undergraduate students during studying. Awareness of the risk of overconsumption of caffeine intake should be implemented in the future. However, this study did not include all types of food choices and drinks. Thus, frequency of eating energy dense food during night studying among students should be conducted in the future.


Asunto(s)
Café , Calidad del Sueño , Humanos , Índice de Masa Corporal , Universidades , Encuestas y Cuestionarios , Estudiantes
3.
J Bioeth Inq ; 2023 Dec 04.
Artículo en Inglés | MEDLINE | ID: mdl-38047997

RESUMEN

In recent years, the genetic testing and selection of IVF embryos, known as preimplantation genetic testing (PGT), has gained much traction in clinical assisted reproduction for preventing transmission of genetic defects. However, a more recent ethically and morally controversial development in PGT is its possible use in selecting IVF embryos for optimal intelligence quotient (IQ) and other non-disease-related socially desirable traits, such as tallness, fair complexion, athletic ability, and eye and hair colour, based on polygenic risk scores (PRS), in what is referred to as PGT-P. Artificial intelligence (AI) and machine learning-based analysis of big data sets collated from genome sequencing of specific human ethnic populations can be used to estimate an individual embryo's likelihood of developing such multifactorial traits by analysing the combination of specific genetic variants within its genome. Superficially, this technique appears compliant with Islamic principles and ethics. Because there is no modification of the human genome, there is no tampering with Allah's creation (taghyir khalq Allah). Nevertheless, a more critical analysis based on the five maxims of Islamic jurisprudence (qawa'id fiqhiyyah) that are often utilized in discourses on Islamic bioethics, namely qasd (intention), yaqin̄ (certainty), darar (injury), darura (necessity), and `urf (custom), would instead reveal some major ethical and moral flaws of this new medical technology in the selection of non-disease-related socially desirable traits, and its non-compliance with the spirit and essence of Islamic law (shariah). Muslim scholars, jurists, doctors, and biomedical scientists should debate this further and issue a fatwa on this new medical technology platform.

4.
Sci Rep ; 13(1): 8014, 2023 05 17.
Artículo en Inglés | MEDLINE | ID: mdl-37198277

RESUMEN

Inflammatory processes are increasingly attributed to macrophage polarization. Proinflammatory macrophages promote T helper (Th) 1 response, tissue repair, and Th2 responses. Detection of macrophages in tissue sections is facilitated by CD68. Our study is focused on the expression of CD68 and the estimation of proinflammatory cytokines in children's patients with chronic tonsillitis secondary to vitamin D supplementation. This hospital-based Randomized prospective case-control study was conducted on 80 children with chronic tonsillitis associated with vitamin D deficiency where (40 received vitamin D 50,000 IU weekly for 3-6 months and 40 received 5 ml distilled water as placebo). The serum 25-hydroxyvitamin D [25(OH)D] was measured using an Enzyme-linked immunosorbent assay on all included children. Different histological and immunohistochemical studies for the detection of CD68 were done. There was a significantly lower serum level of 25(OH)D in the placebo group versus the vitamin D group (P < 0.001). The levels of pro-inflammatory cytokines, TNFα, and IL-2 significantly increased in the placebo group as compared to the vitamin D group (P < 0.001). The increased level of IL-4 and IL-10 in the placebo group as compared to the vitamin D group was insignificant (P = 0.32, 0.82) respectively. Vitamin D supplementation alleviated the deleterious effect of chronic tonsillitis on the histological structure of the tonsil. Tonsillar tissues of the children in the control and vitamin D groups demonstrated a highly statistically significantly lower number of CD68 immunoexpressing cells compared with those in the placebo group (P < 0.001). Low vitamin D may play a role in chronic tonsillitis. Vitamin D supplementation could help reduce the occurrence of chronic tonsillitis in susceptible children.


Asunto(s)
Tonsilitis , Deficiencia de Vitamina D , Niño , Humanos , Estudios de Casos y Controles , Colecalciferol , Citocinas , Suplementos Dietéticos , Vitamina D , Vitaminas
5.
Int J Gen Med ; 15: 5681-5691, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35747780

RESUMEN

Background: COVID-19 (SARS-CoV-2/2019-nCoV) is now a major public health threat to the world. Olfactory dysfunctions (ODs) are considered potential indicating symptoms and early case identification triaging for coronavirus disease 2019 (COVID-19). The most common reported comorbidities are diabetes mellitus, chronic lung disease, and cardiovascular disease. The objective of this study was to evaluate prevalence of different types of smell disorders in patients with laboratory-confirmed COVID-19 infection and impact of involved systemic diseases. Methodology: A cross-sectional retrospective study has been done for patients with laboratory-confirmed COVID-19 infection (mild-to-moderate). The data collected from patient's files and developed online electronic questionnaire (WhatsApp) based on the patients most common and recurrent reported data including: a) symptoms of olfactory dysfunction and associated covid19 symptoms fever and headache, cough, sore throat, pneumonia, nausea, vomiting and diarrhea, arthralgia and myalgia and taste dysfunction. b) Associated systemic diseases including: diabetes, hypertension, asthma, chronic renal disease, chorionic liver disease and hypothyroidism. Results: Of 308 patients confirmed with Covid-19 infection, (72.4%) developed OD distributed as follows; complete anosmia (57.8%), troposmia (8.4%), hyposmia (2.9%), partial anosmia (2.6%) and euosmia (0.6%). Significantly increased prevalence of diabetes, hypertension asthma in the group with olfactory dysfunction (p < 0.001), chronic liver disease (p = 0.005), and hypothyroidism (p = 0.03). Conclusion: The development of ODs after Covid-19 infection was associated with mild disease form and lower hospitalization. In addition, it showed significant relationship with preexisting systemic diseases. Anosmia is the common modality of ODs.

6.
Med J Malaysia ; 77(1): 1-5, 2022 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-35086987

RESUMEN

INTRODUCTION: Endometriosis is a challenging disease to treat, and patients may eventually need in vitro fertilisation with Intracytoplasmic sperm injection (IVF/ICSI) to conceive after other modalities failed. There are inconsistent outcomes of IVF performance in patients with endometriosis especially with highly purified human menotropin gonadotrophin (hMG). This study was commenced to determine whether the use of hMG affects the IVF outcome in different stage of endometriosis. MATERIALS AND METHODS: This is an observational study. Eighty-seven women who had endometriosis confirmed surgically and underwent IVF/ICSI treatment, stimulated with hMG alone were included. Based on the revised American Society for Reproductive Medicine (rASRM), the participants were classified as early endometriosis (I/II) (n=39) or advanced endometriosis (III/IV) (n=35). The main outcome measures used were clinical pregnancy rate. RESULTS: Women with advanced endometriosis had a lower oocyte yield, less good quality day-3 embryos and lower clinical pregnancy rate compared with the mild endometriosis. However, higher fertilisation rate were recorded in advanced stage endometriosis compared to milder disease. CONCLUSIONS: The rASRM classification of endometriosis is valuable in predicting IVF outcome as advanced endometriosis performs poorly compared to a milder disease. Highly purified hMG could be an alternative as an ovarian stimulation in endometriosis.


Asunto(s)
Endometriosis , Inyecciones de Esperma Intracitoplasmáticas , Endometriosis/cirugía , Femenino , Fertilización In Vitro , Humanos , Inducción de la Ovulación , Embarazo , Índice de Embarazo
7.
Sci Rep ; 11(1): 20702, 2021 10 19.
Artículo en Inglés | MEDLINE | ID: mdl-34667216

RESUMEN

Breast cancer is one of the most reported cancers that can lead to death. Despite the advances in diagnosis and treatment procedures, the possibility of cancer recurrences is still high in many cases. With that in consideration, researchers from all over the world are showing interest in the unique features of Graphene oxide (GO), such as its excellent and versatile physicochemical properties, to explore further its potential and benefits towards breast cancer cell treatment. In this study, the cell viability and electrical response of GO, in terms of resistivity and impedance towards the breast cancer cells (MCF7) and normal breast cells (MCF10a), were investigated by varying the pH and concentration of GO. Firstly, the numbers of MCF7 and MCF10a were measured after being treated with GO for 24 and 48 h. Next, the electrical responses of these cells were evaluated by using interdigitated gold electrodes (IDEs) that are connected to an LCR meter. Based on the results obtained, as the pH of GO increased from pH 5 to pH 7, the number of viable MCF7 cells decreased while the number of viable MCF10a slightly increased after the incubation period of 48 h. Similarly, the MCF7 also experienced higher cytotoxicity effects when treated with GO concentrations of more than 25 µg/mL. The findings from the electrical characterization of the cells observed that the number of viable cells has corresponded to the impedance of the cells. The electrical impedance of MCF7 decreased as the number of highly insulating viable cell membranes decreased. But in contrast, the electrical impedance of MCF10a increased as the number of highly insulating viable cell membranes increased. Hence, it can be deduced that the GO with higher pH and concentration influence the MCF7 cancer cell line and MCF10a normal breast cell.


Asunto(s)
Neoplasias de la Mama/tratamiento farmacológico , Supervivencia Celular/efectos de los fármacos , Grafito/farmacología , Apoptosis/efectos de los fármacos , Mama/efectos de los fármacos , Recuento de Células , Línea Celular Tumoral , Impedancia Eléctrica/uso terapéutico , Electrodos , Femenino , Oro/farmacología , Humanos , Células MCF-7 , Recurrencia Local de Neoplasia/tratamiento farmacológico
8.
Med J Malaysia ; 76(3): 417-418, 2021 05.
Artículo en Inglés | MEDLINE | ID: mdl-34031343

RESUMEN

Fertility preservation is significant for oncology patients to maintain their ability to start a family when they are ready. Onco-fertility, as a discipline, exists at the intersection of oncology and reproductive medicine that safeguards and expands the fertility options for cancer survivors, by facilitating early intervention and suitable treatment with favourable outcomes. Successful fertility preservation requires a comprehensive networking among the gynaecologists, oncologists, pathologists, imaging and other specialists, involved in diagnosing and treating cancer in the reproductive age group. There are several ways in which fertility can be preserved, like role of gonadotrophin releasing hormone analogues, in vitro maturation, and cryopreservation.


Asunto(s)
Preservación de la Fertilidad , Neoplasias , Criopreservación , Fertilidad , Humanos , Malasia , Neoplasias/terapia
9.
Int J Anal Chem ; 2018: 8150610, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-29692811

RESUMEN

Rigidoporus microporus, Ganoderma philippii, and Phellinus noxius are root rot rubber diseases and these fungi should be kept under control with environmentally safe compounds from the plant sources. Thus, an antifungal compound isolated from Catharanthus roseus was screened for its effectiveness in controlling the growth of these fungi. The antifungal compound isolated from C. roseus extract was determined through thin layer chromatography (TLC) and nuclear magnetic resonance (NMR) analysis. Each C. roseus of the DCM extracts was marked as CRD1, CRD2, CRD3, CRD4, CRD5, CRD6, and CRD7, respectively. TLC results showed that all of the C. roseus extracts peaked with red colour at Rf = 0.61 at 366 nm wavelength, except for CRD7. The CRD4 extract was found to be the most effective against R. microporus and G. philippii with inhibition zones of 3.5 and 1.9 mm, respectively, compared to that of other extracts. These extracts, however, were not effective against P. noxius. The CRD4 extract contained ursolic acid that was detected by NMR analysis and the compound could be developed as a biocontrol agent for controlling R. microporus and G. philippii. Moreover, little or no research has been done to study the effectiveness of C. roseus in controlling these fungi.

10.
J Hosp Infect ; 54(1): 68-73, 2003 May.
Artículo en Inglés | MEDLINE | ID: mdl-12767850

RESUMEN

This study investigated knowledge about infection control amongst doctors and nurses through a cross-sectional survey conducted between March and May 2001 in three Birmingham, UK teaching hospitals. Seventy-five doctors and 143 nurses, representing 7% and 4%, respectively, of potential respondents, participated in the study measuring knowledge of, attitudes towards, and compliance with universal precautions. Overall knowledge of risks of blood-borne virus (BBV) transmission from an infected patient after needlestick injury was low [44.0% for hepatitis B virus (HBV), 38.1% for hepatitis C virus (HCV), 54.6% for human immunodeficiency virus (HIV)]. There were significant differences between doctors and nurses concerning the estimations of HBV (e-antigen +) (P=0.006) and HIV (P<0.001) transmission risks. Eighty-six percent of nurses stated that they treat each patient as if they are carrying a BBV compared with 41% of doctors. Doctors and nurses differed significantly in their attitudes about and reported compliance with washing hands before and after patient contact and with wearing gloves when taking blood (P<0.001 for all). Doctors consistently de-emphasized the importance of, and reported poor compliance with, these procedures. Doctors were also more likely to state that they re-sheath used needles manually than were nurses (P<0.001). Thirty-seven percent of respondents reported that they had suffered a needlestick injury with a used needle, with doctors more likely to be injured than nurses (P=0.005). Twenty-eight percent of these doctors and 2% of the nurses did not report their needlestick injuries (P=0.004). Education, monitoring, improved availability of resources, and disciplinary measures for poor compliance are necessary to improve infection control in hospitals, especially amongst doctors.


Asunto(s)
Actitud del Personal de Salud , Transmisión de Enfermedad Infecciosa de Paciente a Profesional/prevención & control , Enfermeras y Enfermeros , Exposición Profesional/estadística & datos numéricos , Médicos , Adulto , Competencia Clínica , Estudios Transversales , Inglaterra/epidemiología , Femenino , Adhesión a Directriz , Infecciones por VIH/prevención & control , Hepatitis B/prevención & control , Hepatitis C/prevención & control , Hospitales de Enseñanza , Humanos , Control de Infecciones/métodos , Control de Infecciones/normas , Masculino , Persona de Mediana Edad , Encuestas y Cuestionarios , Precauciones Universales
11.
Cancer Lett ; 104(2): 225-31, 1996 Jul 12.
Artículo en Inglés | MEDLINE | ID: mdl-8665492

RESUMEN

Tumor biopsies (paraffin embedded tissue) obtained from 45 Saudi patients with non-Hodgkin's lymphoma (NHL) were examined for the incidence of p53 mutations screened by polymerase chain reaction (PCR) and single strand conformation polymorphism (SSCP) analysis. DNA sequencing was carried out to confirm the occurrence of p53 mutation in PCR products showing abnormal migration by SSCP analysis. Only 1/45 samples showed the incidence of a homozygous mutation at codon 179 (exon 5) of the p53 gene that replaces histidine with tyrosine. The data showed that the frequency of p53 mutations was very low in Saudi NHL. Our results are consistent with the general observation that the p53 mutation is rather infrequent in hematopoietic malignancies like NHLs.


Asunto(s)
Genes p53 , Linfoma no Hodgkin/genética , Mutación , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Secuencia de Bases , Niño , Preescolar , ADN de Neoplasias/análisis , Exones , Femenino , Humanos , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Polimorfismo Conformacional Retorcido-Simple
12.
Nature ; 369(6476): 156-60, 1994 May 12.
Artículo en Inglés | MEDLINE | ID: mdl-8177321

RESUMEN

The mitogen-activated protein (MAP) kinases Erk-1 and Erk-2 are proline-directed kinases that are themselves activated through concomitant phosphorylation of tyrosine and threonine residues. The kinase p54 (M(r) 54,000), which was first isolated from cycloheximide-treated rats, is proline-directed like Erks-1/2, and requires both Tyr and Ser/Thr phosphorylation for activity. p54 is, however, distinct from Erks-1/2 in its substrate specificity, being unable to phosphorylate pp90rsk but more active in phosphorylating the c-Jun transactivation domain. Molecular cloning of p54 reveals a unique subfamily of extracellularly regulated kinases. Although they are 40-45% identical in sequence to Erks-1/2, unlike Erks-1/2 the p54s are only poorly activated in most cells by mitogens or phorbol esters. However, p54s are the principal c-Jun N-terminal kinases activated by cellular stress and tumour necrosis factor (TNF)-alpha, hence they are designated stress-activated protein kinases, or SAPKs. SAPKs are also activated by sphingomyelinase, which elicits a subset of cellular responses to TNF-alpha (ref. 9). SAPKs therefore define a new TNF-alpha and stress-activated signalling pathway, possibly initiated by sphingomyelin-based second messengers, which regulates the activity of c-Jun.


Asunto(s)
Proteínas Quinasas Dependientes de Calcio-Calmodulina/metabolismo , Proteínas Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-jun/metabolismo , Células 3T3 , Secuencia de Aminoácidos , Animales , Proteínas Quinasas Dependientes de Calcio-Calmodulina/clasificación , Proteínas Quinasas Dependientes de Calcio-Calmodulina/genética , Línea Celular , Clonación Molecular , Cicloheximida/farmacología , Activación Enzimática/efectos de los fármacos , Calor , Ratones , Proteína Quinasa 1 Activada por Mitógenos , Proteína Quinasa 3 Activada por Mitógenos , Proteína Quinasa 9 Activada por Mitógenos , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Datos de Secuencia Molecular , Fosforilación , Proteínas Quinasas/clasificación , Proteínas Quinasas/genética , Proteínas Serina-Treonina Quinasas , Proteínas Tirosina Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-jun/clasificación , Proteínas Proto-Oncogénicas c-jun/genética , Ratas , Homología de Secuencia de Aminoácido , Transducción de Señal , Esfingomielina Fosfodiesterasa/farmacología , Porcinos , Factor de Necrosis Tumoral alfa/farmacología
13.
Thyroid ; 4(2): 151-3, 1994.
Artículo en Inglés | MEDLINE | ID: mdl-7919996

RESUMEN

We have amplified a 285 base pair by nested polymerase chain reaction 38 nucleotide downstream from the most 5' transcription initiation site (7) encoding 55 of the 57 residues of exon 1 of the human TSH receptor. These DNA were amplified from 10 tissue blocks of thyroid tissue removed at subtotal thyroidectomy from 10 patients with Graves' disease. The amplified 285 nucleotide fragments were sequenced in search of mutations in the coding region of exon 1 and polymorphism in the 120 nucleotides of the untranslated region upstream of the first ATG codon. No such variations were found. We conclude that the polymorphism or mutation of the part of the TSH receptor extracellular domain encoded by exon 1 and of sequences immediately upstream of the first ATG codon are not relevant to the pathogenesis of Graves' disease.


Asunto(s)
Exones/genética , Enfermedad de Graves/genética , Receptores de Tirotropina/genética , Glándula Tiroides/química , Secuencia de Bases , Humanos , Datos de Secuencia Molecular , Mutación , Reacción en Cadena de la Polimerasa , Polimorfismo Genético , Biosíntesis de Proteínas
14.
Biochemistry ; 32(30): 7727-38, 1993 Aug 03.
Artículo en Inglés | MEDLINE | ID: mdl-7688567

RESUMEN

Phorbol ester tumor promoters (TPA) activate the endogenous erk/MAP kinases and Rsk S6 kinases but not the p70S6 kinase in COS cells. DNA sequences encoding the rat Rsk-1 S6 kinase (homologous to Xenopus rsk alpha), modified by insertion of a peptide epitope at the polypeptide aminoterminus, were expressed transiently in COS cells. TPA stimulates the 40S and peptide kinase activity of the recombinant epitope-tagged Rsk-1, as well as the extent of Rsk-1 autophosphorylation in vitro (32P-Ser >> 32P-Thr). Indications that the conformation of the recombinant Rsk-1 polypeptide is substantially changed after activation by TPA in situ include a retarded mobility of the Rsk-1 polypeptide on SDS-PAGE and the appearance of new 32P-peptides during autophosphorylation in vitro. All these features of the TPA-activated Rsk-1 S6 kinase are abolished by dephosphorylation of the kinase in vitro with Ser/Thr phosphatase-2A. TPA increases 32P incorporation into recombinant Rsk-1 by 2-3-fold (32P-Ser >> 32P-Thr). Peptide mapping exhibits a single major 32P-peptide in Rsk-1 isolated from unstimulated cells and 10-12 additional 32P peptides after TPA treatment in situ. Phosphorylation of basal or phosphatase-2A-treated recombinant Rsk-1 in vitro with erk2/MAP kinase increases Rsk-1 40S kinase, peptide kinase, and autophosphorylating activity, retards migration of Rsk-1 polypeptides on SDS-PAGE, and generates new sites of Rsk-1 autophosphorylation in vitro. By contrast, TPA-activated Rsk-1 is not altered in these properties by autophosphorylation in vitro. By contrast, TPA-activated Rsk-1 is not altered in these properties by phosphorylation in vitro with erk2/MAP kinase. Activation of Rsk-1 in situ with TPA diminishes by over 90% the extent of Rsk-1 phosphorylation achieved in vitro by erk2/MAP kinase, as compared to the parallel phosphorylation of a phosphatase-2A-treated Rsk-1; basal Rsk-1 is intermediate. Peptide maps of phosphatase-2A-treated Rsk-1 after phosphorylation in vitro with erk2/MAP kinase exhibit 32P-peptides that comigrate with nearly all of the 32P-peptides present in TPA-activated-32P Rsk-1 labeled in situ, plus several 32P-peptides characteristic of Rsk-1 autophosphorylation in vitro.(ABSTRACT TRUNCATED AT 400 WORDS)


Asunto(s)
Quinasas de Proteína Quinasa Activadas por Mitógenos , Proteínas Serina-Treonina Quinasas/metabolismo , Proteínas Tirosina Quinasas/metabolismo , Acetato de Tetradecanoilforbol/farmacología , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Células Cultivadas , ADN , Activación Enzimática , Epítopos , MAP Quinasa Quinasa 1 , Datos de Secuencia Molecular , Fosforilación , Proteínas Serina-Treonina Quinasas/genética , Ratas , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Proteínas Quinasas S6 Ribosómicas , Xenopus
15.
Protein Expr Purif ; 3(3): 236-45, 1992 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-1392620

RESUMEN

Separation of copurifying protease activity from recombinant human Müllerian inhibiting substance (rhMIS) bound to a monoclonal antibody immunoaffinity column by a high-salt wash results in cleaner preparations of rhMIS resistant to cleavage upon storage. In addition, an inhibitor of rhMIS antiproliferative activity is removed. Proteolytic cleavages produced by either a copurifying protease or exogenous plasmin occur at residues 229 and 427 but do not abolish rhMIS biological activity. This report details the modified immunoaffinity column isolation protocol suitable for proteins such as rhMIS and describes the biochemical and antiproliferative properties of this protein.


Asunto(s)
Cromatografía de Afinidad , Glicoproteínas , Inhibidores de Crecimiento/aislamiento & purificación , Técnicas de Inmunoadsorción , Proteínas Recombinantes de Fusión/aislamiento & purificación , Hormonas Testiculares/aislamiento & purificación , Secuencia de Aminoácidos , Animales , Hormona Antimülleriana , Anticuerpos Monoclonales/inmunología , Western Blotting , Células CHO , Bovinos , División Celular/efectos de los fármacos , Cricetinae , Endopeptidasas/metabolismo , Inhibidores de Crecimiento/inmunología , Inhibidores de Crecimiento/farmacología , Humanos , Concentración de Iones de Hidrógeno , Datos de Secuencia Molecular , Fragmentos de Péptidos/inmunología , Proteínas Recombinantes de Fusión/inmunología , Proteínas Recombinantes de Fusión/farmacología , Temperatura , Hormonas Testiculares/inmunología , Hormonas Testiculares/farmacología , Células Tumorales Cultivadas/efectos de los fármacos
16.
Proc Natl Acad Sci U S A ; 87(21): 8550-4, 1990 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-2236064

RESUMEN

The molecular structure of a rat hepatoma 70-kDa insulin/mitogen-stimulated S6 protein kinase, obtained by molecular cloning, is compared to that of a rat homolog of the 85-kDa Xenopus S6 protein kinase alpha; both kinases were cloned from H4 hepatoma cDNA libraries. The 70-kDa S6 kinase (calculated molecular mass of 59,186 Da) exhibits a single catalytic domain that is most closely related in amino acid sequence (56% identity) to the amino-terminal, kinase C-like domain of the rat p85 S6 kinase (calculated molecular mass of 82,695 Da); strong similarity extends through a further 67 residues carboxyl-terminal to the catalytic domain (40% identity), corresponding to a region also conserved among the kinase C family. Outside of this segment of approximately 330 amino acids, the structures of the p70 and p85 S6 kinases diverge substantially. The p70 S6 kinase is known to be activated through serine/threonine phosphorylation by unidentified insulin/mitogen-activated protein kinases. A model for the regulation of p70 S6 protein kinase activity is proposed wherein the low activity of the unphosphorylated enzyme results from the binding of a basic, inhibitory pseudosubstrate site (located carboxyl-terminal to the extended catalytic domain) to an acidic substrate binding region (located amino-terminal to the catalytic domain); substrate binding is thereby prevented. S6 kinase activation requires displacement of this inhibitory segment, which is proposed to occur consequent to its multiple phosphorylation. The putative autoinhibitory segment contains several serine and threonine residues, each followed directly by a proline residue. This motif may prevent autophosphorylation but permit transphosphorylation; two of these serine residues reside in a maturation promoting factor (MPF)/cdc-2 consensus motif. Thus, hormonal regulation of S6 kinase may involve the action of MPF/cdc-2 or protein kinases with related substrate specificity.


Asunto(s)
Insulina/farmacología , Neoplasias Hepáticas Experimentales/enzimología , Mitógenos/farmacología , Proteínas Quinasas/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Cicloheximida/farmacología , Activación Enzimática , Biblioteca de Genes , Hígado/efectos de los fármacos , Hígado/enzimología , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Proteínas Quinasas/aislamiento & purificación , Proteínas Quinasas/metabolismo , ARN Neoplásico/genética , ARN Neoplásico/aislamiento & purificación , Ratas , Proteínas Quinasas S6 Ribosómicas , Homología de Secuencia de Ácido Nucleico , Xenopus
18.
Blood ; 74(6): 1888-93, 1989 Nov 01.
Artículo en Inglés | MEDLINE | ID: mdl-2679910

RESUMEN

Circulating IgG autoantibodies that produce cytoplasmic immunofluorescence staining of ethanol-fixed normal neutrophils have recently been found in a large percentage of patients with active Wegener's granulomatosis. Such autoantibodies are rarely found in other diseases and are therefore virtually diagnostic of Wegener's granulomatosis. The nature of the neutrophil antigen defined by these autoantibodies is controversial and the roles of the antigen and/or autoantibodies in the pathogenesis of Wegener's granulomatosis are unknown. We studied serum samples that produce the cytoplasmic pattern of staining from 10 patients with a diagnosis of Wegener's granulomatosis. By Western blot analysis, all 10 sera reacted with a 29-Kd neutrophil protein (p29). We generated a mouse monoclonal antibody directed against this antigen. The monoclonal antibody produced the same immunofluorescence staining pattern as the serum autoantibodies and was used to affinity-purify p29. The purified antigen had a novel N-terminal sequence homologous to members of the serine proteinase family and bound to radiolabeled diisopropyl fluorophosphate (DFP). We conclude that the neutrophil antigen responsible for the cytoplasmic staining pattern produced by autoantibodies in patients with active Wegener's granulomatosis is a distinctive serine proteinase.


Asunto(s)
Autoantígenos/fisiología , Enfermedades Autoinmunes/inmunología , Granulomatosis con Poliangitis/inmunología , Neutrófilos/enzimología , Serina Endopeptidasas/inmunología , Secuencia de Aminoácidos , Anticuerpos Monoclonales/inmunología , Autoanticuerpos/inmunología , Western Blotting , Citoplasma/enzimología , Citoplasma/inmunología , Técnica del Anticuerpo Fluorescente , Granulomatosis con Poliangitis/enzimología , Humanos , Datos de Secuencia Molecular , Peso Molecular , Neutrófilos/inmunología
20.
Eur J Biochem ; 161(1): 1-6, 1986 Nov 17.
Artículo en Inglés | MEDLINE | ID: mdl-3096729

RESUMEN

Antibodies prepared against a homogeneous preparation of Co-eIF-2A20 [Ahmad et al. (1985) J. Biol. Chem. 260, 6955-6959] reacted with several polypeptides including an 80-kDa polypeptide present in a crude yeast ribosomal salt wash. This 80-kDa polypeptide, containing Co-eIF-2A (Co-eIF-2A80) activity, has been extensively purified using a two-step purification procedure involving an immunoaffinity column chromatograph prepared using antibodies against Co-eIF-2A20 (fraction II) and hydroxyapatite chromatography (fraction III). The factors, eIF-2 + homogeneous Co-eIF-2A80 (fraction III) promoted Met-tRNAf.40S complex formation with an AUG codon but not with a physiological mRNA or a polyribonucleotide messenger poly(U,G) whereas eIF-2 + a partially purified Co-eIF-2A80 preparation (fraction II) promoted Met-tRNAf.40S complex formation with an AUG codon as well as with globin mRNA and poly(U,G) messenger. This factor-promoted Met-tRNAf binding to 40S ribosomes depends absolutely on the presence of a polyribonucleotide messenger containing an initiation codon (such as AUG or GUG). Other polyribonucleotide messengers tested, such as poly(U), poly(A) and poly(A,C) were completely ineffective in this binding reaction. This result indicates that the Met-tRNAf.40S.mRNA complex is formed by a direct interaction between Met-tRNAf, 40S ribosomes and the initiation site in mRNA. A mechanism has been proposed for Met-tRNAf.40S.mRNA complex formation in yeast.


Asunto(s)
Proteínas Portadoras/aislamiento & purificación , Proteínas Fúngicas/biosíntesis , Factores de Iniciación de Péptidos/aislamiento & purificación , ARN Mensajero/metabolismo , Aminoacil-ARN de Transferencia/metabolismo , Proteínas Ribosómicas/biosíntesis , Proteínas Ribosómicas/metabolismo , Saccharomyces cerevisiae/metabolismo , Proteínas Portadoras/fisiología , Factores de Iniciación de Péptidos/fisiología , Péptidos/aislamiento & purificación , Péptidos/fisiología , ARN Mensajero/fisiología , Proteínas de Unión al ARN
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